Immunohistochemistry analysis of Troponin I Rabbit Recombinant Oligoclonal Antibody (710580) was done on paraffin embedded mouse heart tissue sections. To expose target proteins, heat induced epitope retrieval was performed using Tris-EDTA (pH 9.0) buffer for 15 minutes. Following antigen retrieval, tissues were blocked in 0.2% BSA with 0.1% cold fish skin gelatin in 1X PBS for 1 hour in a humidified chamber. The tissues were then probed at a dilution of 1:100 with ABfinity™ Troponin I Recombinant Rabbit Oligoclonal Antibody (710580) or blocking buffer alone as negative control for 3 hours at room temperature in a humidified chamber. Detection was performed using Alexa Fluor® 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes in a humidified chamber. Tissues were counterstained with SlowFade® Gold Antifade Mountant with DAPI (S36938). The images were captured using a Nikon microscope at 20X magnification.<br />
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||human Troponin I|
|Contains||0.09% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||Assay Dependent|
|Immunofluorescence (IF)||Assay Dependent|
|Immunohistochemistry (IHC)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Troponin and tropomyosin are the main proteins responsible for calcium regulation of actin-myosin interactions. Tropomyosin is a rod-shaped molecule that binds in the groove of the actin helix, and prevents the attachment of the myosin fiber heads. The troponin complex consists of three polypeptide subunits, troponin T, troponin I, and troponin C. Troponin T (for tropomyosin binding unit) attaches the troponin complex to tropomyosin which results in tropomyosin being situated in the myosin-binding site on the actin molecule. Troponin I (for inhibitory subunit) binds to actin and troponin T, and this complex inhibits actin-myosin binding even in the presence of calcium. Troponin C (for calcium-binding subunit) binds to troponin T and troponin I, and makes the complex dependent upon calcium. With calcium present, troponin C binds up to 4 molecules and changes the position of the troponin and tropomyosin complex, so that myosin is able to bind to the actin filament. Troponin I is present in three different isoforms in mammals, a slow-twitch skeletal muscle-specific form, a fast-twitch muscle-specific form, and a cardiac muscle-specific form.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.